Composts of poultry litter or dairy manure differentially affect survival of enteric bacteria in fields with spinach.

AIMS: The aim was to determine the survival and persistence of Escherichia coli in soil amended with compost from different manure sources. METHOD AND RESULTS: Complex interactions of abiotic and biotic factors on E. coli survival were characterized in field experiment plots receiving randomly assigned compost treatments: dairy windrow, dairy vermicompost, poultry windrow or no compost. Biomass, activity and function of indigenous microbial communities in the composts and soils were measured concurrently to determine whether mechanisms of compost were driven by biotic or abiotic properties. E. coli persisted in compost containing poultry amendments but not in composts containing dairy or no amendments. Poultry compost contained more NH4 -N and a distinct microbial community compared to dairy and no compost treatments. A laboratory experiment performed on compost extracts suggested that E. coli survived better in extracts devoid of indigenous microbes as long as bioavailable nutrients were plentiful. CONCLUSIONS: Dairy-based composts are less likely to support E. coli survival than poultry-based composts. SIGNIFICANCE AND IMPACT OF THE STUDY: Results aid in risk assessment of the use of different types of manure-based compost and soil amendments in fruit and vegetable production by elucidating the roles of nutrient and microbial community composition on survival of E. coli in amended field soils.

Use of zero-valent iron biosand filters to reduce Escherichia coli O157:H12 in irrigation water applied to spinach plants in a field setting.

AIMS: Zero-valent iron (ZVI) filters may provide an efficient method to mitigate the contamination of produce crops through irrigation water. METHODS: A field-scale system was utilized to evaluate the effectiveness of a biosand filter (S), a biosand filter with ZVI incorporated (ZVI) and a control (C, no treatment) in decontaminating irrigation water. An inoculum of c.8·5log CFU100ml(-1) of Escherichia coli O157:H12 was introduced to all three column treatments in 20-l doses. Filtered waters were subsequently overhead irrigated to 'Tyee' spinach plants. Water, spinach plant and soil samples were obtained on days 0, 1, 4, 6, 8, 10, 13 and 15 and analysed for E. coli O157:H12 populations. RESULTS: ZVI filters inactivated c.6logCFU100ml(-1) E. coli O157:H12 during filtration on day 0, significantly (P<0·05) more than S filter (0·49CFU100ml(-1)) when compared to control on day 0 (8·3log CFU100ml(-1)). On day 0, spinach plants irrigated with ZVI-filtered water had significantly lower E. coli O157 counts (0·13logCFUg(-1)) than spinach irrigated with either S-filtered (4·37logCFUg(-1)) or control (5·23logCFUg(-1)) water. Soils irrigated with ZVI-filtered water contained E. coli O157:H12 populations below the detection limit (2logCFUg(-1)), while those irrigated with S-filtered water (3·56logCFUg(-1)) were significantly lower than those irrigated with control (4·64logCFUg(-1)). CONCLUSIONS: ZVI biosand filters were more effective in reducing E. coli O157:H12 populations in irrigation water than sand filters. SIGNIFICANCE AND IMPACT OF THE STUDY: Zero-valent ion treatment may be a cost-effective mitigation step to help small farmers reduce risk of foodborne E. coli infections associated with contamination of leafy greens.

Enhanced solid-liquid separation of dairy manure with natural flocculants.

The aim of this study was to determine the effectiveness of natural flocculants to reduce solids and nutrient loads in dairy cow wastewater using solid-liquid separation; chitosan was used as a model. Its use efficiency and optimum application rate were determined using flushed dairy cow manure of varied strengths - 0.4%, 0.8%, 1.6%, and 3.2% total solids (TS) content. Treatments consisted of nine rates of chitosan. The flocculated manure was dewatered using 1-mm and 0.25-mm screens. Separation by screening alone was not effective; average efficiencies were about 60% for total suspended solids (TSS), 22% for total Kjeldahl nitrogen (TKN), and 26% for total phosphorus (TP). Mixing with chitosan before screening substantially increased separation. At optimum chitosan rate (0.5g/L for the highest strength effluent), separation efficiencies were >95% for TSS, >73% for TKN, and >54% for TP. The results of this study indicate that natural flocculants such as chitosan are useful for the solid-liquid separation treatment of livestock wastewater.

Potential of olive mill waste and compost as biobased pesticides against weeds, fungi, and nematodes.

The phytotoxic and antimicrobial properties of olive mill wastes have been widely investigated and demonstrated over the past decade. However, their potential utilization as biodegradable pesticides against plant pathogens is still poorly understood. In this study, a series of laboratory bioassays was designed to test the inhibitory effects of sterile water extracts of two-phase olive mill waste (TPOMW) and TPOMW composts with different degrees of stabilization on several different plant pathogens. Fungicidal properties of TPOMW extracts, assayed in a microwell assay format, showed that the growth of Phytophthora capsici was consistently and strongly inhibited by all TPOMW extracts diluted 1:10 (w:v). In contrast, suppression of Pythium ultimum and Botrytis cinerea by the extracts was not as strong and depended on the specific TPOMW sample. Mature compost inhibited P. capsici and B. cinerea at dilutions as great as 1:50, w:v. Neither TPOMW nor TPOMW compost extracts were able to inhibit the growth of the basidiomycete root rot agent Rhizoctonia solani. In addition, studies were conducted on the allelopathic effects of TPOMW extracts on seed germination of four highly invasive and globally distributed weeds (Amaranthus retroflexus, Solanum nigrum, Chenopodium album and Sorghum halepense). Both the TPOMW and immature TPOMW compost extracts substantially inhibited germination of A. retroflexus and S. nigrum, whereas mature composts extracts only partially reduced the germination of S. nigrum. Finally, TPOMW extracts strongly inhibited egg hatch and second-stage juvenile (J2) motility of the root-knot nematode Meloidogyne incognita. However, only higher concentrations of stage-one and stage-two TPOMW compost extracts exerted a suppressive effect on both J2 motility and on egg hatch. The study shows the high potential of naturally occurring chemicals present in TPOMW and TPOMW composts that should be further investigated as bio-pesticides for their use in sustainable agricultural systems.

Factors affecting compost tea as a potential source of Escherichia coli and Salmonella on fresh produce.

Compost tea (CT) is an unheated on-farm infusion of compost used as a spray or soil drench to promote plant growth and control foliar and root diseases. Because food safety involves all aspects from farm to fork, CT should meet basic microbiological criteria for water quality. This report describes the effects of two CT production processes, aerated and nonaerated, on growth and survival of foodborne pathogens and fecal coliforms. Seven commercially available nutrients used to supplement CT were tested individually and in combination for their effects on the growth of Escherichia coli and Salmonella. Compost containing 10(1) to 10(3) CFU/g initial concentrations of E. coli O157:H7 and Salmonella Enteritidis were used to assess growth and survival responses to aerated CT (36-h preparations) and nonaerated CT (8.5-day preparations). Pathogen and fecal coliform populations were undetectable by 8.5 days in nonaerated CT without nutrient supplements. E. coli O157:H7 decreased to below detection levels in aerated CT at 36 h without the use of supplements. In contrast, the addition of commercially formulated mixtures or combinations of nutrient supplements resulted in growth of E. coli O157: H7, Salmonella, and fecal coliforms by 1 to 4 log CFU/g in both aerated and nonaerated CT. When nutrient supplements were added, aerated CT sustained higher concentrations of E. coli O157:H7, Salmonella, and fecal coliforms than did nonaerated CT. Thus, addition of supplements supports growth of human pathogens from very low initial concentrations in both aerated and nonaerated CT and should be avoided when CT is used on fresh produce.

On-farm and postharvest processing sources of bacterial contamination to melon rinds.

Multistate and international foodborne illness outbreaks, particularly involving cantaloupe and often involving rare Salmonella spp., have increased dramatically over the past 13 years. This study assessed the sources and extent of melon rind contamination in production fields and at processing and packing facilities. In the spring of 1999, cantaloupe (Cucumis melo L. [reticulatus group] cv. Cruiser) sampled from two sites in the Rio Grande River Valley showed that postharvest-processed melon rinds often had greater plate counts of bacterial contaminants than field-fresh melons. Cantaloupe in the field had 2.5 to 3.5 log CFU g(-1) rind total coliforms by aerobic plate counts, whereas washed melons had 4.0 to 5.0 log CFU g(-1). In the fall of 1999, coliforms on honeydew melons (C. melo [inodorous group] cv. Honey Brew) ranged from 2.6 to 3.7 log CFU g(-1) after processing, and total and fecal coliforms and enterococci never fell below 2.5 log CFU g(-1). A hydrocooler at another site contaminated cantaloupe rinds with up to 3.4 log CFU g(-1) total and fecal enterococci; a secondary rinse with chlorinated water incompletely removed these bacteria. Sources of coliforms and enterococci were at high levels in melon production soils, especially in furrows that were flood irrigated, in standing water at one field, and in irrigation water at both sites. At one processing facility, wash water pumped from the Rio Grande River may not have been sufficiently disinfected prior to use. Because soil, irrigation water, and process water were potential sources of bacterial contamination, monitoring and management on-farm and at processing and packing facilities should focus on water quality as an important control point for growers and packers to reduce bacterial contamination on melon rinds.

Taxon-specific oligonucleotide primers for detection of two ancient endomycorrhizal fungi, Glomus occultum and Glomus brasilianum.

A unique oligonucleotide pair, GOCC56:GOCC427, was designed that correctly primed specific amplification of a approximately 370-bp sequence spanning the ITS and 5.8S rDNA regions of Glomus occultum and Glomus brasilianum. In addition, this primer pair successfully detected G. occultum and G. brasilianum DNA in nested PCR using a primary PCR product amplified from highly diluted extracts of colonized corn (Zea mays) roots using modified ITS1:ITS4 primers. A second primer pair, GBRAS86:GBRAS388, primed specific amplification of a approximately 200-bp sequence spanning the ITS and 5.8S rDNA regions present only in G. brasilianum and Glomus strain GR582. Combined use of both primer pairs provides the means to detect and differentiate two ancient endomycorrhizal species, G. occultum and G. brasilianum, undetectable by standard root staining procedures. Sequence analysis showed that the purported G. occultum strain GR582 is likely a strain of G. brasilianum.

Microbially mediated growth suppression and death of salmonella in composted sewage sludge.

The role of compost microflora in the suppression of salmonella regrowth in composted sewage sludge was investigated. Microbial inhibition studies of salmonella growth were conducted on nutrient agar, in composts that had been subjected to different temperatures in compost piles, and in radiation sterilized composts inoculated with selected fractions of the compost microflora. Agar assays of inhibition indicated that bacteria and actinomycetes were not suppressive to salmonellae, but a few fungi were. However, compost inoculation assays showed consistently that fungi were not suppressive, but bacteria and actinomycetes were. In compost inoculation assays, microbial antagonists, when present, either killed salmonellae or reduced their growth rate. No suppression of salmonellae occurred in compost taken from 70°C compost-pile zones despite the presence and growth of many types of microbes. With greater numbers and kinds of microbes in 55°C compost, salmonella growth was suppressed 100-10,000-fold. Salmonellae died when inoculated into compost from unheated zones (25-40°C) of piles. Prior colonization of compost with only noncoliform gram-negative bacteria suppressed salmonellae growth 3,000-fold. Coliforms when inoculated prior to salmonellae accounted for 75% of salmonella die-off. Mesophilic curing to allow colonization of curing piles in their entirety by gram-negative bacteria, especially coliforms, should be an effective way to prevent repopulation by salmonellae.

Acute bronchoconstriction induced by cotton dust: dose-related responses to endotoxin and other dust factors.

Fifty-four healthy humans, selected for their acute airway responsiveness to cotton dust, had spirometric tests immediately before and after 6 hours of exposure to card-generated cotton dust from seven different cottons (of several grades and growing regions). During exposures, we measured airborne concentrations of viable fungi and bacteria (total and gram negative), vertically elutriated gravimetric dust, and vertically elutriated endotoxin. Correlation between each of these five exposure indices and exposure-related acute changes in forced expiratory volume in 1 s showed a statistically significant relationship between all of the indices except concentration of viable fungi. Of the other four indices, endotoxin was the most highly correlated (r = -0.94; p less than 0.00001), and gravimetric dust was the least correlated (r = -0.34; p less than 0.05). These findings suggest that gram-negative endotoxin may play a major role in the acute pulmonary response to inhaled cotton dust.

Bacteria on closed-boll and commercially harvested cotton.

The bacterial content of specially treated cottons used by other investigators to test human pulmonary responses to cotton dust was examined. Cotton from Lubbock, Tex. and Stoneville, Miss. were either (i) harvested by machine and handled as commercial bale cotton, (ii) harvested as closed bolls with bracts intact and opened under special conditions, (iii) harvested as closed bolls, with bracts being removed and opened under special conditions, or (iv) harvested by (stoneville only). Bacillus spp. were isolated from all samples and predominated in cotton from Stoneville. Enterobacter agglomerans was isolated from all but one sample, the Stoneville closed-boll bract-removed cotton, and predominated in Lubbock samples. Aerogenic and anaerogenic biogroups of E. agglomerans were isolated; only aerogenic strain b of E. agglomerans was present in samples from both locations. Klebsiella ozaenae and K. pneumoniae were isolated only from Lubbock samples. Cotton from Lubbock yielded 100 to 1,000 times more bacteria, both total and gram negative, than did comparably treated cotton from Stoneville. Thus, differences in growing and processing conditions at the two locations were associated with large differences in the bacterial content of the cotton, but harvesting green bolls and removing bracts had little effect. The bacterial content of Stoneville washed cotton, and it paralleled the differences reported (Boehlecke et al., Am. Rev. Respir, Dis. 123:152, 1981) in pulmonary function responses when subjects were exposed to dust (0.6 mg/m3) from these two cottons. Levels of gram-negative and total bacteria on all samples were comparable to those previously reported for field-weathered cottons from various locations throughout the world.

Dispersal of Aspergillus fumigatus from Sewage Sludge Compost Piles Subjected to Mechanical Agitation in Open Air.

Aerosolization of the thermophilous fungal opportunist Aspergillus fumigatus from mechanically agitated compost piles was examined at a pilot-scale sewage sludge composting facility and two other selected test sites. Aerosols of A. fumigatus downwind from stationary compost piles were insignificant in comparison with those downwind from agitated piles. These aerosols were generated by a front-end loader moving and dropping compost. Aerial concentrations of the fungus at distances downwind from the point of emission were used to determine an emission rate for A. fumigatus associated with the moving operations. The maximum emission rate, 4.6 x 10A. fumigatus particles per s, was used to calculate predicted concentrations in an unobstructed plume with restrictive, neutral, and dispersive atmospheric mixing conditions up to 1 km downwind from the emission source.

A guide to the recent literature on aspergillosis as caused by Aspergillus fumigatus, a fungus frequently found in self-heating organic matter.

Spores of Aspergillus fumigatus have been found to be abundantly present in the outdoor air at a site where large scale experimental composting of sewage sludge is in progress at Beltsville, Maryland. The health significance of this finding, for that site and for others in the future, is still only incompletely understood. Further studies are in progress to characterize absolute concentrations of the spores of the fungus in air at the site, spore dispersal by air from composting operations, and background environmental spore levels in air. The present paper contains a list of references to papers on health effects of A. fumigatus, many published in the past ten years, along with a review of the same designed to assist the reader in finding information on particular aspects of the subject in the literature. It is intended primarily as an aid to individuals interested in sludge composting and wishing to attain an insight into the A. fumigatus-composting situation, but it may also interest others concerned with other substrates which become moldy at 40--50 C. A. fumigatus has been found in great numbers in naturally and artificially heated environments such as decaying leaves, compost heaps, solar heated sloughs, cooling canals for nuclear power generators, silos, grain storage bins, boiler rooms, detritus around steam turbines and sauna baths. The evident practical merits of sludge composting have been described elsewhere; the information presented here has its main significance in respect to requirements for choice of locations for composting sites and to process and design criteria.

Occurrence of Aspergillus fumigatus during composting of sewage sludge.

Aspergillus fumigatus, a medically important fungal opportunist and respiratory allergen, was isolated from woodchips and sewage sludge used in the production of compost at the U.S. Department of Agriculture's composting research facility in Beltsville, Md. It was also regularly isolated as a dominant fungus during forced aeration composting and after 30 days in an unaerated stationary curing pile; in both cases, the fungus was found in pile zones with temperatures less than 60 degrees C. Compost stored outdoors in stationary unaerated piles from 1 to 4 months after screening out of woodchips contained easily detectable amounts of A. fumigatus in the exterior pile zones (0- to 25-cm depths). Semiquantitative studies of the airspora at the composting site revealed that A. fumigatus constituted 75% of the total viable mycoflora captured. At locations 320 m to 8 km from the compost site, the fungus constituted only 2% of the total viable mycoflora in the air. Of 21 samples of commercially available potting soil, one had levels of A. fumigatus nearly equivalent to those of 1-month-old storage compost; 15 others had lower but detectable levels.